Статті в журналах: "NRT2"

1

Zoghbi-Rodríguez, Normig M., Samuel David Gamboa-Tuz, Alejandro Pereira-Santana, Luis C. Rodríguez-Zapata, Lorenzo Felipe Sánchez-Teyer, and Ileana Echevarría-Machado. "Phylogenomic and Microsynteny Analysis Provides Evidence of Genome Arrangements of High-Affinity Nitrate Transporter Gene Families of Plants." International Journal of Molecular Sciences 22, no. 23 (December 3, 2021): 13036. http://dx.doi.org/10.3390/ijms222313036.

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Nitrate transporter 2 (NRT2) and NRT3 or nitrate-assimilation-related 2 (NAR2) proteins families form a two-component, high-affinity nitrate transport system, which is essential for the acquisition of nitrate from soils with low N availability. An extensive phylogenomic analysis across land plants for these families has not been performed. In this study, we performed a microsynteny and orthology analysis on the NRT2 and NRT3 genes families across 132 plants (Sensu lato) to decipher their evolutionary history. We identified significant differences in the number of sequences per taxonomic group and different genomic contexts within the NRT2 family that might have contributed to N acquisition by the plants. We hypothesized that the greater losses of NRT2 sequences correlate with specialized ecological adaptations, such as aquatic, epiphytic, and carnivory lifestyles. We also detected expansion on the NRT2 family in specific lineages that could be a source of key innovations for colonizing contrasting niches in N availability. Microsyntenic analysis on NRT3 family showed a deep conservation on land plants, suggesting a high evolutionary constraint to preserve their function. Our study provides novel information that could be used as guide for functional characterization of these gene families across plant lineages.

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2

Zhang, Jingying, Zhijun Han, Yue Lu, Yanfei Zhao, Yaping Wang, Jiayue Zhang, Haoran Ma, and Yu Zhu Han. "Genome-wide identification, structural and gene expression analysis of the nitrate transporters (NRTs) family in potato (Solanum tuberosum L.)." PLOS ONE 16, no. 10 (October 21, 2021): e0257383. http://dx.doi.org/10.1371/journal.pone.0257383.

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Nitrogen (N2) is the most important source of mineral N for plant growth, which was mainly transported by nitrate transporters (NRTs). However, little is known about the NRT gene family in potato. In this study, StNRT gene family members were identified in potato. In addition, we performed StNRT subfamily classification, gene structure and distribution analysis, and conserved domain prediction using various bioinformatics tools. Totally, 39 StNRT gene members were identified in potato genome, including 33, 4 and 2 member belong to NRT1, NRT2, and NRT3, respectively. These 39 StNRT genes were randomly distributed on all chromosomes. The collinearity results show that StNRT members in potato are closely related to Solanum lycopersicum and Solanum melongena. For the expression, different members of StNRT play different roles in leaves and roots. Especially under sufficient nitrogen conditions, different members have a clear distribution in different tissues. These results provide valuable information for identifying the members of the StNRT family in potato and could provide functional characterization of StNRT genes in further research.

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3

Newstead, Simon, and Joanne Parker. "Crystal structure of the plant nitrate transporter NRT1.1." Acta Crystallographica Section A Foundations and Advances 70, a1 (August 5, 2014): C1487. http://dx.doi.org/10.1107/s205327331408512x.

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Nitrogen uptake and assimilation is a key limiting factor for plant growth and crop productivity and also acts a major signaling molecule, controlling many aspects of plant development. Many plants obtain nitrogen through the uptake of nitrate from the soil via specific membrane transporters. Two families of nitrate transporter have been identified that act within the root cell, termed NRT1 and NRT2. NRT1 members are predominantly low affinity transporters, with KM values in the millimolar range, whereas NRT2 members are high affinity transporters, with KM values in the low micromolar range. Dual affinity transporter systems are used in biology to allow the cell to respond to changes in an external nutrient supply. In the case of nitrate transport in plants, decreasing levels of external nitrate cause an increase in the expression of NRT2 family transporter genes, in particular NRT2.1, allowing the cell to take up more of the available nitrate. However, plants have evolved a faster way of responding to nitrate levels involving post-translational control of nitrate uptake. NRT1.1 also known as CHL1, is a dual affinity nitrate transporter. In response to decreasing levels of nitrate, NRT1.1 is capable of switching between low and high KM modes, a switch achieved through the post-translational phosphorylation of an intracellular threonine. Here I will present our recently determined crystal structures of NRT1.1 in both the apo and nitrate bound forms. Together with in vitro binding and transport data we identify key residues involved in nitrate recognition and provide the first biochemical explanation for the phosphorylation controlled `dual affinity' switch observed in NRT1.1. Finally we present our model for the molecular basis of nitrate uptake via this transporter.

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4

CRISCUOLO, GIUSEPPINA, VLADIMIR TOTEV VALKOV, AURORA PARLATI, LUDOVICO MARTIN ALVES, and MAURIZIO CHIURAZZI. "Molecular characterization of the Lotus japonicus NRT1(PTR) and NRT2 families." Plant, Cell & Environment 35, no. 9 (April 19, 2012): 1567–81. http://dx.doi.org/10.1111/j.1365-3040.2012.02510.x.

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5

Wang, Xiaoli, Xiaofeng Cai, Chenxi Xu, and Quanhua Wang. "Identification and characterization of the NPF, NRT2 and NRT3 in spinach." Plant Physiology and Biochemistry 158 (January 2021): 297–307. http://dx.doi.org/10.1016/j.plaphy.2020.11.017.

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6

Ruffel, Sandrine, Valentin Chaput, Jonathan Przybyla-Toscano, Ian Fayos, Catalina Ibarra, Tomas Moyano, Cécile Fizames, et al. "Genome-wide analysis in response to nitrogen and carbon identifies regulators for root AtNRT2 transporters." Plant Physiology 186, no. 1 (February 5, 2021): 696–714. http://dx.doi.org/10.1093/plphys/kiab047.

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Abstract In Arabidopsis (Arabidopsis thaliana), the High-Affinity Transport System (HATS) for root nitrate (NO3−) uptake depends mainly on four NRT2 NO3− transporters, namely NRT2.1, NRT2.2, NRT2.4, and NRT2.5. The HATS is the target of many regulations to coordinate nitrogen (N) acquisition with the N status of the plant and with carbon (C) assimilation through photosynthesis. At the molecular level, C and N signaling pathways control gene expression of the NRT2 transporters. Although several regulators of these transporters have been identified in response to either N or C signals, the response of NRT2 gene expression to the interaction of these signals has never been specifically investigated, and the underlying molecular mechanisms remain largely unknown. To address this question we used an original systems biology approach to model a regulatory gene network targeting NRT2.1, NRT2.2, NRT2.4, and NRT2.5 in response to N/C signals. Our systems analysis of the data identified three transcription factors, TGA3, MYC1, and bHLH093. Functional analysis of mutants combined with yeast one-hybrid experiments confirmed that all three transcription factors are regulators of NRT2.4 or NRT2.5 in response to N or C signals. These results reveal a role for TGA3, MYC1, and bHLH093 in controlling the expression of root NRT2 transporter genes.

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7

Okamoto, Mamoru, J. John Vidmar, and Anthony D. M. Glass. "Regulation of NRT1 and NRT2 Gene Families of Arabidopsis thaliana: Responses to Nitrate Provision." Plant and Cell Physiology 44, no. 3 (March 15, 2003): 304–17. http://dx.doi.org/10.1093/pcp/pcg036.

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8

Guo, Tiancai, Hongmei Xuan, Yingying Yang, Lina Wang, Liting Wei, Yonghua Wang, and Guozhang Kang. "Transcription Analysis of Genes Encoding the Wheat Root Transporter NRT1 and NRT2 Families During Nitrogen Starvation." Journal of Plant Growth Regulation 33, no. 4 (June 19, 2014): 837–48. http://dx.doi.org/10.1007/s00344-014-9435-z.

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9

Miller, A., X. Fan, Q. Shen, and S. Smith. "Expression and functional analysis of rice NRT2 nitrate transporters." Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 146, no. 4 (April 2007): S241. http://dx.doi.org/10.1016/j.cbpa.2007.01.558.

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10

Quesada, A., J. Hidalgo, and E. Fernández. "Three Nrt2 genes are differentially regulated in Chlamydomonas reinhardtii." Molecular and General Genetics MGG 258, no. 4 (May 1998): 373–77. http://dx.doi.org/10.1007/s004380050743.

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11

Slot, Jason C., Kelly N. Hallstrom, Patrick B. Matheny, and David S. Hibbett. "Diversification of NRT2 and the Origin of Its Fungal Homolog." Molecular Biology and Evolution 24, no. 8 (May 19, 2007): 1731–43. http://dx.doi.org/10.1093/molbev/msm098.

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12

Sakamoto, Toshio, Kaori Inoue-Sakamoto, and Donald A. Bryant. "A Novel Nitrate/Nitrite Permease in the Marine CyanobacteriumSynechococcus sp. Strain PCC 7002." Journal of Bacteriology 181, no. 23 (December 1, 1999): 7363–72. http://dx.doi.org/10.1128/jb.181.23.7363-7372.1999.

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ABSTRACT The nrtP and narB genes, encoding nitrate/nitrite permease and nitrate reductase, respectively, were isolated from the marine cyanobacterium Synechococcus sp. strain PCC 7002 and characterized. NrtP is a member of the major facilitator superfamily and is unrelated to the ATP-binding cassette-type nitrate transporters that previously have been described for freshwater strains of cyanobacteria. However, NrtP is similar to the NRT2-type nitrate transporters found in diverse organisms. An nrtP mutant strain consumes nitrate at a 4.5-fold-lower rate than the wild type, and this mutant grew exponentially on a medium containing 12 mM nitrate at a rate approximately 2-fold lower than that of the wild type. The nrtP mutant cells could not consume nitrite as rapidly as the wild type at pH 10, suggesting that NrtP also functions in nitrite uptake. A narB mutant was unable to grow on a medium containing nitrate as a nitrogen source, although this mutant could grow on media containing urea or nitrite with rates similar to those of the wild type. Exogenously added nitrite enhanced the in vivo activity of nitrite reductase in the narBmutant; this suggests that nitrite acts as a positive effector of nitrite reductase. Transcripts of the nrtP andnarB genes were detected in cells grown on nitrate but were not detected in cells grown on urea or ammonia. Transcription of thenrtP and narB genes is probably controlled by the NtcA transcription factor for global nitrogen control. The discovery of a nitrate/nitrite permease in Synechococcussp. strain PCC 7002 suggests that significant differences in nutrient transporters may occur in marine and freshwater cyanobacteria.

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13

Du, Run-Jie, Ze-Xuan Wu, Zhao-Xi Yu, Peng-Feng Li, Jian-Yu Mu, Jie Zhou, Jia-Na Li, and Hai Du. "Genome-Wide Characterization of High-Affinity Nitrate Transporter 2 (NRT2) Gene Family in Brassica napus." International Journal of Molecular Sciences 23, no. 9 (April 29, 2022): 4965. http://dx.doi.org/10.3390/ijms23094965.

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Nitrate transporter 2 (NRT2) plays an essential role in Nitrogen (N) uptake, transport, utilization, and stress resistance. In this study, the NRT2 gene family in two sequenced Brassica napus ecotypes were identified, including 31 genes in ‘Zhongshuang11’ (BnaZSNRT2s) and 19 in ‘Darmor-bzh’ (BnaDarNRT2s). The candidate genes were divided into three groups (Group I−III) based on phylogenetic analyses, supported by a conserved intron-exon structure in each group. Collinearity analysis revealed that the large expansion of BnaZSNRT2s attributed to allopolyploidization of ancestors Brassica rapa and Brassica oleracea, and small-scale duplication events in B. napus. Transcription factor (TF) binding site prediction, cis-element analysis, and microRNA prediction suggested that the expressions of BnaZSNRT2s are regulated by multiple factors, and the regulatory pattern is relatively conserved in each group and is tightly connected between groups. Expression assay showed the diverse and differentiated spatial-temporal expression profiles of BnaZSNRT2s in Group I, but conserved patterns were observed in Group II/III; and the low nitrogen (LN) stress up-regulated expression profiles were presented in Group I−III, based on RNA-seq data. RT-qPCR analyses confirmed that BnaZSNRT2.5A-1 and BnaZSNRT2.5C-1 in Group II were highly up-regulated under LN stress in B. napus roots. Our results offer valid information and candidates for further functional BnaZSNRT2s studies.

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14

Luo, Bingbing, Man Xu, Limei Zhao, Peng Xie, Yi Chen, Wendy Harwood, Guohua Xu, Xiaorong Fan, and Anthony J. Miller. "Overexpression of the High-Affinity Nitrate Transporter OsNRT2.3b Driven by Different Promoters in Barley Improves Yield and Nutrient Uptake Balance." International Journal of Molecular Sciences 21, no. 4 (February 15, 2020): 1320. http://dx.doi.org/10.3390/ijms21041320.

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Improving nitrogen use efficiency (NUE) is very important for crops throughout the world. Rice mainly utilizes ammonium as an N source, but it also has four NRT2 genes involved in nitrate transport. The OsNRT2.3b transporter is important for maintaining cellular pH under mixed N supplies. Overexpression of this transporter driven by a ubiquitin promoter in rice greatly improved yield and NUE. This strategy for improving the NUE of crops may also be important for other cereals such as wheat and barley, which also face the challenges of nutrient uptake balance. To test this idea, we constructed transgenic barley lines overexpressing OsNRT2.3b. These transgenic barley lines overexpressing the rice transporter exhibited improved growth, yield, and NUE. We demonstrated that NRT2 family members and the partner protein HvNAR2.3 were also up-regulated by nitrate treatment (0.2 mM) in the transgenic lines. This suggests that the expression of OsNRT2.3b and other HvNRT2 family members were all up-regulated in the transgenic barley to increase the efficiency of N uptake and usage. We also compared the ubiquitin (Ubi) and a phloem-specific (RSs1) promoter-driven expression of OsNRT2.3b. The Ubi promoter failed to improve nutrient uptake balance, whereas the RSs1 promoter succeed in increasing the N, P, and Fe uptake balance. The nutrient uptake enhancement did not include Mn and Mg. Surprisingly, we found that the choice of promoter influenced the barley phenotype, not only increasing NUE and grain yield, but also improving nutrient uptake balance.

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15

Higuera, Jose, Victoria Calatrava, Zaira González, Vicente Mariscal, Jose Siverio, Emilio Fernández, and Aurora Galván. "NRT2.4 and NRT2.5 Are Two Half-Size Transporters from the Chlamydomonas NRT2 Family." Agronomy 6, no. 1 (March 19, 2016): 20. http://dx.doi.org/10.3390/agronomy6010020.

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16

Orsel, Mathilde, Anne Krapp, and Françoise Daniel-Vedele. "Analysis of the NRT2 Nitrate Transporter Family in Arabidopsis. Structure and Gene Expression." Plant Physiology 129, no. 2 (May 24, 2002): 886–96. http://dx.doi.org/10.1104/pp.005280.

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17

Filleur, Sophie, Marie-France Dorbe, Miguel Cerezo, Mathilde Orsel, Fabienne Granier, Alain Gojon, and Françoise Daniel-Vedele. "An Arabidopsis T-DNA mutant affected in Nrt2 genes is impaired in nitrate uptake." FEBS Letters 489, no. 2-3 (January 31, 2001): 220–24. http://dx.doi.org/10.1016/s0014-5793(01)02096-8.

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18

Liu, Xiaojing, Yajiao Zhao, and Feng Hao. "Development of nitrogen efficiency screening system in alfalfa (Medicago sativa L.) and analysis of alfalfa nitrogen efficiency types." PeerJ 10 (May 6, 2022): e13343. http://dx.doi.org/10.7717/peerj.13343.

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Screening high nitrogen (N) efficiency crops is crucial to utilize resources rationally and reduce N losses. In this research, the biomass, morphological and N-related parameters of 28 alfalfa (Medicago sativa L.) cultivars were assessed at seedling stage. Then, we selected representative materials to compare the changes in stem-leaf dry weight (SDW), total root length (RL) and plant N accumulation (PNA) during whole period. Lastly, we analyzed the expressions of NRT2 and AMT1 genes of alfalfa cultivars. The correlation coefficients between SDW, PDW, RL, RV, SNA, RNA, and PNA were all in the range of 0.522∼0.996. The coefficient of variations of SDW, PDW, RL, RV, SNA and PNA were all more than 20% under low and medium N levels. Though the comprehensive evaluation and cluster analysis, the comprehensive value of LW6010, Gannong NO.5, Longmu 806, Giant 2, Giant 601, Zhaodong, Crown were greater than 0.5 under low and medium N levels; the comprehensive value of Gannong NO.3, Gannong NO.4, Xinjiangdaye, Xinmu NO.1 were less than 0.5 under low N level, but were greater than 0.5 under medium N level. The comprehensive value of Gannong NO.7 Gannong NO.9, Longmu 801, Gongnong NO.3, Elite, Sadie 10, Giant 551 were greater than 0.5 under low N level, but were lesser than 0.5 under medium N level; and those of Longdong, Gannong NO.8, Gongnong NO.1, Reindee, Goldqueen, Weston, Tourists, Giant 6, Algonquin, Sadie 7 were lesser than 0.5 under low and medium N levels. Four N efficiency types of alfalfa cultivars were classified: (1) Very efficient; (2) Efficient; (3) Anti-efficient; and (4) Inefficient.The SDW, RL and PNA of LW6010 were higher than Longdong in each growth period. The expressions of NRT2 and AMT1 genes were highest for LW6010, and lowest for Longdong. So, N efficiency parameters assessed at seedling stage include: SDW, PDW, RL, RV, SNA and PNA. We developed new classification system of N efficiency types of alfalfa cultivars. It proved its effectiveness on 28 alfalfa in China.

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19

Nakamura, Y., Y. Umemiya, K. Masuda, H. Inoue, and M. Fukumoto. "Molecular cloning and expression analysis of cDNAs encoding a putative Nrt2 nitrate transporter from peach." Tree Physiology 27, no. 4 (April 1, 2007): 503–10. http://dx.doi.org/10.1093/treephys/27.4.503.

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20

Swapnil, Prashant, Mukesh Meena, and Ashwani K. Rai. "Molecular interaction of nitrate transporter proteins with recombinant glycinebetaine results in efficient nitrate uptake in the cyanobacterium Anabaena PCC 7120." PLOS ONE 16, no. 11 (November 18, 2021): e0257870. http://dx.doi.org/10.1371/journal.pone.0257870.

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Nitrate transport in cyanobacteria is mediated by ABC-transporter, which consists of a highly conserved ATP binding cassette (ABC) and a less conserved transmembrane domain (TMD). Under salt stress, recombinant glycinebetaine (GB) not only protected the rate of nitrate transport in transgenic Anabaena PCC 7120, rather stimulated the rate by interacting with the ABC-transporter proteins. In silico analyses revealed that nrtA protein consisted of 427 amino acids, the majority of which were hydrophobic and contained a Tat (twin-arginine translocation) signal profile of 34 amino acids (1–34). The nrtC subunit of 657 amino acids contained two hydrophobic distinct domains; the N-terminal (5–228 amino acids), which was 59% identical to nrtD (the ATP-binding subunit) and the C-terminal (268–591), 28.2% identical to nrtA, suggesting C-terminal as a solute binding domain and N-terminal as ATP binding domain. Subunit nrtD consisted of 277 amino acids and its N-terminal (21–254) was an ATP binding motif. Phylogenetic analysis revealed that nitrate-ABC-transporter proteins are highly conserved among the cyanobacterial species, though variation existed in sequences resulting in several subclades. Nostoc PCC 7120 was very close to Anabaena variabilis ATCC 29413, Anabaena sp. 4–3 and Anabaena sp. CA = ATCC 33047. On the other, Nostoc spp. NIES-3756 and PCC 7524 were often found in the same subclade suggesting more work before referring it to Anabaena PCC 7120 or Nostoc PCC 7120. The molecular interaction of nitrate with nrtA was hydrophilic, while hydrophobic with nrtC and nrtD. GB interaction with nrtACD was hydrophobic and showed higher affinity compared to nitrate.

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21

Wang, Jiang, Norman Hüner, and Lining Tian. "Identification and molecular characterization of the Brachypodium distachyon NRT2 family, with a major role of BdNRT2.1." Physiologia Plantarum 165, no. 3 (April 30, 2018): 498–510. http://dx.doi.org/10.1111/ppl.12716.

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22

Maeda, Shin-ichi, Risa Aoba, Yuma Nishino, and Tatsuo Omata. "A Novel Bacterial Nitrate Transporter Composed of Small Transmembrane Proteins." Plant and Cell Physiology 60, no. 10 (June 14, 2019): 2180–92. http://dx.doi.org/10.1093/pcp/pcz112.

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AbstractA putative silent gene of the freshwater cyanobacterium Synechococcus elongatus strain PCC 7942, encoding a small protein with two transmembrane helices, was named nrtS, since its overexpression from an inducible promoter conferred nitrate uptake activity on the nitrate transport-less NA4 mutant of S. elongatus. Homologs of nrtS, encoding proteins of 67–118 amino acid residues, are present in a limited number of eubacteria including mostly cyanobacteria and proteobacteria, but some others, e.g. the actinobacteria of the Mycobacterium tuberculosis complex, also have the gene. When expressed in NA4, the nrtS homolog of the γ-proteobacterium Marinomonas mediterranea took up nitrate with higher affinity for the substrate as compared with the S. elongatus NrtS (Km of 0.49 mM vs. 2.5 mM). Among the 61 bacterial species carrying the nrtS homolog, the marine cyanobacterium Synechococcus sp. strain PCC 7002 is unique in having two nrtS genes (nrtS1 and nrtS2) located in tandem on the chromosome. Coexpression of the two genes in NA4 resulted in nitrate uptake with a Km (NO3−) of 0.15 mM, while expression of either of the two resulted in low-affinity nitrate uptake activity with Km values of >3 mM, indicating that NrtS1 and NrtS2 form a heteromeric transporter complex. The heteromeric transporter was shown to transport nitrite as well. A Synechococcus sp. strain PCC 7002 mutant defective in the nitrate transporter (NrtP) showed a residual activity of nitrate uptake, which was ascribed to the NrtS proteins. Blue-native PAGE and immunoblotting analysis suggested a hexameric structure for the NrtS proteins.

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23

Pechkovskaya, S. A., N. A. Knyazev, O. V. Matantseva, A. K. Emelyanov, I. V. Telesh, S. O. Skarlato, and N. A. Filatova. "Dur3 and nrt2 genes in the bloom-forming dinoflagellate Prorocentrum minimum: Transcriptional responses to available nitrogen sources." Chemosphere 241 (February 2020): 125083. http://dx.doi.org/10.1016/j.chemosphere.2019.125083.

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24

Willmann, Anita, Stefanie Thomfohrde, Robert Haensch, and Uwe Nehls. "The poplar NRT2 gene family of high affinity nitrate importers: Impact of nitrogen nutrition and ectomycorrhiza formation." Environmental and Experimental Botany 108 (December 2014): 79–88. http://dx.doi.org/10.1016/j.envexpbot.2014.02.003.

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25

Krapp, Anne, Vincent Fraisier, Wolf-Rudiger Scheible, Alberto Quesada, Alain Gojon, Mark Stitt, Michel Caboche, and Francoise Daniel-Vedele. "Expression studies of Nrt2:1Np, a putative high-affinity nitrate transporter: evidence for its role in nitrate uptake." Plant Journal 14, no. 6 (June 1998): 723–31. http://dx.doi.org/10.1046/j.1365-313x.1998.00181.x.

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26

Kemppainen, Minna J., and Alejandro G. Pardo. "LbNrtRNA silencing in the mycorrhizal symbiontLaccaria bicolorreveals a nitrate-independent regulatory role for a eukaryotic NRT2-type nitrate transporter." Environmental Microbiology Reports 5, no. 3 (January 25, 2013): 353–66. http://dx.doi.org/10.1111/1758-2229.12029.

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27

Kechid, Maya, Guilhem Desbrosses, Lydia Gamet, Loren Castaings, Fabrice Varoquaux, Abdelhamid Djekoun, and Bruno Touraine. "Arabidopsis Growth-Promotion and Root Architecture Responses to the Beneficial Rhizobacterium Phyllobacterium brassicacearum Strain STM196 Are Independent of the Nitrate Assimilatory Pathway." Plants 11, no. 1 (January 4, 2022): 128. http://dx.doi.org/10.3390/plants11010128.

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Phyllobacterium brassicacearum STM196, a plant growth-promoting rhizobacterium isolated from roots of oilseed rape, stimulates Arabidopsis growth. We have previously shown that the NRT2.5 and NRT2.6 genes are required for this growth promotion response. Since these genes are members of the NRT2 family of nitrate transporters, the nitrogen assimilatory pathway could be involved in growth promotion by STM196. We address this hypothesis using two nitrate reductase mutants, G5 deleted in the major nitrate reductase gene NIA2 and G′4-3 altered in both NIA1 and NIA2 genes. Both mutants had a reduced growth rate and STM196 failed to increase their biomass production on a medium containing NO3− as the sole nitrogen source. However, they both displayed similar growth promotion by STM196 when grown on an NH4+ medium. STM196 was able to stimulate lateral roots development of the mutants under both nutrition conditions. Altogether, our results indicate that the nitrate assimilatory metabolism is not a primary target of STM196 interaction and is not involved in the root developmental response. The NIA1 transcript level was reduced in the shoots of nrt2.5 and nrt2.6 mutants suggesting a role for this nitrate reductase isoform independently from its role in nitrate assimilation.

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28

Kang, Lee-Kuo, Hsuan-Fan Wang, and Jeng Chang. "Diversity of Phytoplankton Nitrate Transporter Sequences from Isolated Single Cells and Mixed Samples from the East China Sea and mRNA Quantification." Applied and Environmental Microbiology 77, no. 1 (November 12, 2010): 122–30. http://dx.doi.org/10.1128/aem.01315-10.

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ABSTRACTThe transcript abundances of nitrate transporter genes (Nrt2) were proposed as potential markers for nitrogen deficiency in marine diatoms. To correctly quantify diatomNrt2mRNA in the East China Sea (ECS), we utilized both mixed-species sequencing and single-cell PCR to expand the sequence database for this region. Using the single-cell method of PCR, 9 new diatomNrt2sequences belonging to 5 genera, theNrt2sequences of which have never been reported before, were obtained. On the other hand, 291 sequences homologous toNrt2were retrieved from mixed-species sequencing using degenerate primers, and these sequences were clustered into 12 major groups according to a phylogenetic analysis. Based on sequence alignments, 11 pairs of group-specific PCR primers were designed to detectNrt2mRNA levels in the ECS, and 3 of these primer pairs showed high specificity to target species. In ECS phytoplankton samples, environmental RNA was amplified via antisense RNA amplification followed by cDNA production. Subsequently,Nrt2transcript levels were readily detected using quantitative PCR. Our results indicated that investigating sequence diversity followed by careful primer design and evaluation is a good strategy to quantify the expression of genes of ecologically important phytoplankton.

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Gu, Benguo, Yi Chen, Fang Xie, Jeremy D. Murray, and Anthony J. Miller. "Inorganic Nitrogen Transport and Assimilation in Pea (Pisum sativum)." Genes 13, no. 1 (January 17, 2022): 158. http://dx.doi.org/10.3390/genes13010158.

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The genome sequences of several legume species are now available allowing the comparison of the nitrogen (N) transporter inventories with non-legume species. A survey of the genes encoding inorganic N transporters and the sensing and assimilatory families in pea, revealed similar numbers of genes encoding the primary N assimilatory enzymes to those in other types of plants. Interestingly, we find that pea and Medicago truncatula have fewer members of the NRT2 nitrate transporter family. We suggest that this difference may result from a decreased dependency on soil nitrate acquisition, as legumes have the capacity to derive N from a symbiotic relationship with diazotrophs. Comparison with M. truncatula, indicates that only one of three NRT2s in pea is likely to be functional, possibly indicating less N uptake before nodule formation and N-fixation starts. Pea seeds are large, containing generous amounts of N-rich storage proteins providing a reserve that helps seedling establishment and this may also explain why fewer high affinity nitrate transporters are required. The capacity for nitrate accumulation in the vacuole is another component of assimilation, as it can provide a storage reservoir that supplies the plant when soil N is depleted. Comparing published pea tissue nitrate concentrations with other plants, we find that there is less accumulation of nitrate, even in non-nodulated plants, and that suggests a lower capacity for vacuolar storage. The long-distance transported form of organic N in the phloem is known to be specialized in legumes, with increased amounts of organic N molecules transported, like ureides, allantoin, asparagine and amides in pea. We suggest that, in general, the lower tissue and phloem nitrate levels compared with non-legumes may also result in less requirement for high affinity nitrate transporters. The pattern of N transporter and assimilatory enzyme distribution in pea is discussed and compared with non-legumes with the aim of identifying future breeding targets.

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30

You, Lili, Yu Wang, Tingting Zhang, Yunfeng Zhu, Ning Ren, Xingyu Jiang, and Yang Zhou. "Genome-wide identification of nitrate transporter 2 (NRT2) gene family and functional analysis of MeNRT2.2 in cassava (Manihot esculenta Crantz)." Gene 809 (January 2022): 146038. http://dx.doi.org/10.1016/j.gene.2021.146038.

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31

Charrier, Aurélie, Jean-Baptiste Bérard, Gaël Bougaran, Grégory Carrier, Ewa Lukomska, Nathalie Schreiber, Flora Fournier, et al. "High-affinity nitrate/nitrite transporter genes (Nrt2) inTisochrysis lutea: identification and expression analyses reveal some interesting specificities of Haptophyta microalgae." Physiologia Plantarum 154, no. 4 (February 27, 2015): 572–90. http://dx.doi.org/10.1111/ppl.12330.

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32

Xia, Xinyao, Qiuhe Wei, Chunxia Xiao, Yiping Ye, Zhiqiang Li, Cécile Marivingt-Mounir, Jean-François Chollet, Wende Liu, and Hanxiang Wu. "Genomic survey of NPF and NRT2 transporter gene families in five inbred maize lines and their responses to pathogens infection." Genomics 115, no. 2 (March 2023): 110555. http://dx.doi.org/10.1016/j.ygeno.2022.110555.

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33

Jallouli, Salma, Sawsen Ayadi, Simone Landi, Giorgia Capasso, Giorgia Santini, Zoubeir Chamekh, Inés Zouari, Fatma Ezzahra Ben Azaiez, Youssef Trifa, and Sergio Esposito. "Physiological and Molecular Osmotic Stress Responses in Three Durum Wheat (Triticum Turgidum ssp Durum) Genotypes." Agronomy 9, no. 9 (September 13, 2019): 550. http://dx.doi.org/10.3390/agronomy9090550.

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This study aims to investigate the activities and expression of enzymes of primary metabolism and relate these data with the growth performance of three different durum wheat genotypes (Maali; YT13; and ON66) under osmotic stress. Growth traits—including plant height, dry weight (DW) and relative water content (RWC)—were measured to classify genotypes depending on their tolerance to stress. Several enzymes were investigated: Ascorbate peroxidase (APX), Glutamine Synthetase (GS), Glutamine dehydrogenase (GDH), Glutamate synthase (GOGAT), Glucose 6-phosphate dehydrogenase (G6PDH), and Phosphoenolpyruvate Carboxylase (PEPC). The expression of the cytosolic and plastidic glutamine synthetase (TaGS1 and TaGS2), high affinity nitrate transporters (TaNRT2.3) and Glutamate dehydrogenase (TaGDH) were also detected by qRT-PCR. The results indicated different growth performances among genotypes, indicating Maali and YT13 as tolerant genotypes and ON66 as a drought-susceptible variety. Data showed a decrease in PEPC and increase in APX activities under osmotic stress; a slight decrease in GS activity was observed, together with an increase in G6PDH in all genotypes; GS and NRT2 expressions changed in a similar pattern in the different genotypes. Interestingly, Maali and YT13 showed higher transcript abundance for GDH under stress compared to ON66, suggesting the implication of GDH in protective phenomena upon osmotic stress.

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Rékangalt, David, Régis Pépin, Marie-Christine Verner, Jean-Claude Debaud, Roland Marmeisse, and Laurence Fraissinet-Tachet. "Expression of the nitrate transporter nrt2 gene from the symbiotic basidiomycete Hebeloma cylindrosporum is affected by host plant and carbon sources." Mycorrhiza 19, no. 3 (January 6, 2009): 143–48. http://dx.doi.org/10.1007/s00572-008-0221-2.

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35

Rubio, Lourdes, Jordi Díaz-García, Vítor Amorim-Silva, Alberto P. Macho, Miguel A. Botella, and José A. Fernández. "Molecular Characterization of ZosmaNRT2, the Putative Sodium Dependent High-Affinity Nitrate Transporter of Zostera marina L." International Journal of Molecular Sciences 20, no. 15 (July 26, 2019): 3650. http://dx.doi.org/10.3390/ijms20153650.

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One of the most important adaptations of seagrasses during sea colonization was the capacity to grow at the low micromolar nitrate concentrations present in the sea. In contrast to terrestrial plants that use H+ symporters for high-affinity NO3− uptake, seagrasses such as Zostera marina L. use a Na+-dependent high-affinity nitrate transporter. Interestingly, in the Z. marina genome, only one gene (Zosma70g00300.1; NRT2.1) is annotated to this function. Analysis of this sequence predicts the presence of 12 transmembrane domains, including the MFS domains of the NNP transporter family and the “nitrate signature” that appears in all members of the NNP family. Phylogenetic analysis shows that this sequence is more related to NRT2.5 than to NRT2.1, sharing a common ancestor with both monocot and dicot plants. Heterologous expression of ZosmaNRT2-GFP together with the high-affinity nitrate transporter accessory protein ZosmaNAR2 (Zosma63g00220.1) in Nicotiana benthamiana leaves displayed four-fold higher fluorescence intensity than single expression of ZosmaNRT2-GFP suggesting the stabilization of NRT2 by NAR2. ZosmaNRT2-GFP signal was present on the Hechtian-strands in the plasmolyzed cells, pointing that ZosmaNRT2 is localized on the plasma membrane and that would be stabilized by ZosmaNAR2. Taken together, these results suggest that Zosma70g00300.1 would encode a high-affinity nitrate transporter located at the plasma membrane, equivalent to NRT2.5 transporters. These molecular data, together with our previous electrophysiological results support that ZosmaNRT2 would have evolved to use Na+ as a driving ion, which might be an essential adaptation of seagrasses to colonize marine environments.

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Montanini, Barbara, Arturo R. Viscomi, Angelo Bolchi, Yusé Martin, José M. Siverio, Raffaella Balestrini, Paola Bonfante, and Simone Ottonello. "Functional properties and differential mode of regulation of the nitrate transporter from a plant symbiotic ascomycete." Biochemical Journal 394, no. 1 (January 27, 2006): 125–34. http://dx.doi.org/10.1042/bj20051199.

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Nitrogen assimilation by plant symbiotic fungi plays a central role in the mutualistic interaction established by these organisms, as well as in nitrogen flux in a variety of soils. In the present study, we report on the functional properties, structural organization and distinctive mode of regulation of TbNrt2 (Tuber borchii NRT2 family transporter), the nitrate transporter of the mycorrhizal ascomycete T. borchii. As revealed by experiments conducted in a nitrate-uptake-defective mutant of the yeast Hansenula polymorpha, TbNrt2 is a high-affinity transporter (Km=4.7 μM nitrate) that is bispecific for nitrate and nitrite. It is expressed in free-living mycelia and in mycorrhizae, where it preferentially accumulates in the plasma membrane of root-contacting hyphae. The TbNrt2 mRNA, which is transcribed from a single-copy gene clustered with the nitrate reductase gene in the T. borchii genome, was specifically up-regulated following transfer of mycelia to nitrate- (or nitrite)-containing medium. However, at variance with the strict nitrate-dependent induction commonly observed in other organisms, TbNrt2 was also up-regulated (at both the mRNA and the protein level) following transfer to a nitrogen-free medium. This unusual mode of regulation differs from that of the adjacent nitrate reductase gene, which was expressed at basal levels under nitrogen deprivation conditions and required nitrate for induction. The functional and expression properties, described in the present study, delineate TbNrt2 as a versatile transporter that may be especially suited to cope with the fluctuating (and often low) mineral nitrogen concentrations found in most natural, especially forest, soils.

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Huang, Wan-Ru, Pin-Yi Liu, Jie Hsu, Xiuzhen Li, and Liping Deng. "Assessment of Near-Real-Time Satellite Precipitation Products from GSMaP in Monitoring Rainfall Variations over Taiwan." Remote Sensing 13, no. 2 (January 8, 2021): 202. http://dx.doi.org/10.3390/rs13020202.

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This study assessed four near-real-time satellite precipitation products (NRT SPPs) of Global Satellite Mapping of Precipitation (GSMaP)—NRT v6 (hereafter NRT6), NRT v7 (hereafter NRT7), Gauge-NRT v6 (hereafter GNRT6), and Gauge-NRT v7 (hereafter GNRT7)— in representing the daily and monthly rainfall variations over Taiwan, an island with complex terrain. The GNRT products are the gauge-adjusted version of NRT products. Evaluations for warm (May–October) and cold months (November–April) were conducted from May 2017 to April 2020. By using observations from more than 400 surface gauges in Taiwan as a reference, our evaluations showed that GNRT products had a greater error than NRT products in underestimating the monthly mean rainfall, especially during the warm months. Among SPPs, NRT7 performed best in quantitative monthly mean rainfall estimation; however, when examining the daily scale, GNRT6 and GNRT7 were superior, particularly for monitoring stronger (i.e., more intense) rainfall events during warm and cold months, respectively. Spatially, the major improvement from NRT6 to GNRT6 (from NRT7 to GNRT7) in monitoring stronger rainfall events over southwestern Taiwan was revealed during warm (cold) months. From NRT6 to NRT7, the improvement in daily rainfall estimation primarily occurred over southwestern and northwestern Taiwan during the warm and cold months, respectively. Possible explanations for the differences between the ability of SPPs are attributed to the algorithms used in SPPs. These findings highlight that different NRT SPPs of GSMaP should be used for studying or monitoring the rainfall variations over Taiwan for different purposes (e.g., warning of floods in different seasons, studying monthly or daily precipitation features in different seasons, etc.).

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Huang, Wan-Ru, Pin-Yi Liu, Jie Hsu, Xiuzhen Li, and Liping Deng. "Assessment of Near-Real-Time Satellite Precipitation Products from GSMaP in Monitoring Rainfall Variations over Taiwan." Remote Sensing 13, no. 2 (January 8, 2021): 202. http://dx.doi.org/10.3390/rs13020202.

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Анотація:

This study assessed four near-real-time satellite precipitation products (NRT SPPs) of Global Satellite Mapping of Precipitation (GSMaP)—NRT v6 (hereafter NRT6), NRT v7 (hereafter NRT7), Gauge-NRT v6 (hereafter GNRT6), and Gauge-NRT v7 (hereafter GNRT7)— in representing the daily and monthly rainfall variations over Taiwan, an island with complex terrain. The GNRT products are the gauge-adjusted version of NRT products. Evaluations for warm (May–October) and cold months (November–April) were conducted from May 2017 to April 2020. By using observations from more than 400 surface gauges in Taiwan as a reference, our evaluations showed that GNRT products had a greater error than NRT products in underestimating the monthly mean rainfall, especially during the warm months. Among SPPs, NRT7 performed best in quantitative monthly mean rainfall estimation; however, when examining the daily scale, GNRT6 and GNRT7 were superior, particularly for monitoring stronger (i.e., more intense) rainfall events during warm and cold months, respectively. Spatially, the major improvement from NRT6 to GNRT6 (from NRT7 to GNRT7) in monitoring stronger rainfall events over southwestern Taiwan was revealed during warm (cold) months. From NRT6 to NRT7, the improvement in daily rainfall estimation primarily occurred over southwestern and northwestern Taiwan during the warm and cold months, respectively. Possible explanations for the differences between the ability of SPPs are attributed to the algorithms used in SPPs. These findings highlight that different NRT SPPs of GSMaP should be used for studying or monitoring the rainfall variations over Taiwan for different purposes (e.g., warning of floods in different seasons, studying monthly or daily precipitation features in different seasons, etc.).

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Li, Mengjiao, Hui Tian, and Yajun Gao. "A genome‐wide analysis of NPF and NRT2 transporter gene families in bread wheat provides new insights into the distribution, function, regulation and evolution of nitrate transporters." Plant and Soil 465, no. 1-2 (May 8, 2021): 47–63. http://dx.doi.org/10.1007/s11104-021-04927-8.

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40

Akbudak, M. Aydın, Ertugrul Filiz, and Durmuş Çetin. "Genome-wide identification and characterization of high-affinity nitrate transporter 2 (NRT2) gene family in tomato (Solanum lycopersicum) and their transcriptional responses to drought and salinity stresses." Journal of Plant Physiology 272 (May 2022): 153684. http://dx.doi.org/10.1016/j.jplph.2022.153684.

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Pii, Youry, Massimiliano Alessandrini, Katia Guardini, Anita Zamboni, and Zeno Varanini. "Induction of high-affinity NO3– uptake in grapevine roots is an active process correlated to the expression of specific members of the NRT2 and plasma membrane H+-ATPase gene families." Functional Plant Biology 41, no. 4 (2014): 353. http://dx.doi.org/10.1071/fp13227.

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The phenomenon of NO3– induction in plant roots has been characterised both in herbaceous and woody plants. Grapevine (Vitis vinifera L.) plants, hydroponically grown, showed an increase in NO3– uptake rate in response to anion treatment for different periods in the nutrient solution after 1 week of NO3– deprivation. The expression profile of the two high-affinity NO3– transporters VvNRT2.4A and VvNRT2.4B, and the gene encoding the accessory protein VvNAR2.2 exhibits a similar trend to that of the anion uptake. The induction, also involving the increase in activity and protein levels of plasma membrane H+-ATPase, is correlated with the expression profile of two (VvHA2 and VvHA4) out of eight putative plasma membrane H+-ATPase genes identified in grapevine genome.

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Akhtar, Naureen, Eugenia Karabika, Duncan A. Rouch, James R. Kinghorn, Anthony D. M. Glass, and Shiela E. Unkles. "High-affinity nitrate/nitrite transporters NrtA and NrtB of Aspergillus nidulans exhibit high specificity and different inhibitor sensitivity." Microbiology 161, no. 7 (July 1, 2015): 1435–46. http://dx.doi.org/10.1099/mic.0.000088.

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Zhao, Zeqi, Mengdi Li, Weiwei Xu, Ji-Hong Liu, and Chunlong Li. "Genome-Wide Identification of NRT Gene Family and Expression Analysis of Nitrate Transporters in Response to Salt Stress in Poncirus trifoliata." Genes 13, no. 7 (June 22, 2022): 1115. http://dx.doi.org/10.3390/genes13071115.

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The uptake and transportation of nitrate play a crucial role in plant growth and development. These processes mostly depend on nitrate transporters (NRT), which guarantee the supplement of nutrition in the plant. In this study, genes encoding NRT with Major Facilitator Superfamily (MFS) domain were identified in trifoliate orange (Poncirus trifoliata (L.) Raf.). Totally, 56 NRT1s, 6 NRT2s, and 2 NAR2s were explored. The bioinformation analysis, including protein characteristics, conserved domain, motif, phylogenetic relationship, cis-acting element, and synteny correlation, indicated the evolutionary conservation and functional diversity of NRT genes. Additionally, expression profiles of PtrNRTs in different tissues demonstrated that NRT genes possessed spatio-temporal expression specificity. Further, the salt condition was certified to induce the expression of some NRT members, like PtrNPF2.1, PtrNPF7.4, and PtrNAR2.1, proposing the potential role of these NRTs in salt stress response. The identification of NRT genes and the expression pattern analysis in various tissues and salt stress lay a foundation for future research between nitrogen transport and salt resistance in P. trifoliata.

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Cullinan, Sara B., John D. Gordan, Jianping Jin, J. Wade Harper, and J. Alan Diehl. "The Keap1-BTB Protein Is an Adaptor That Bridges Nrf2 to a Cul3-Based E3 Ligase: Oxidative Stress Sensing by a Cul3-Keap1 Ligase." Molecular and Cellular Biology 24, no. 19 (October 1, 2004): 8477–86. http://dx.doi.org/10.1128/mcb.24.19.8477-8486.2004.

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ABSTRACT The Nrf2 transcription factor promotes survival following cellular insults that trigger oxidative damage. Nrf2 activity is opposed by the BTB/POZ domain protein Keap1. Keap1 is proposed to regulate Nrf2 activity strictly through its capacity to inhibit Nrf2 nuclear import. Recent work suggests that inhibition of Nrf2 may also depend upon ubiquitin-mediated proteolysis. To address the contribution of Keap1-dependent sequestration versus Nrf2 proteolysis, we identified the E3 ligase that regulates Nrf2 ubiquitination. We demonstrate that Keap1 is not solely a cytosolic anchor; rather, Keap1 is an adaptor that bridges Nrf2 to Cul3. We demonstrate that Cul3-Keap1 complexes regulate Nrf2 polyubiquitination both in vitro and in vivo. Inhibition of either Keap1 or Cul3 increases Nrf2 nuclear accumulation, leading to promiscuous activation of Nrf2-dependent gene expression. Our data demonstrate that Keap1 restrains Nrf2 activity via its capacity to target Nrf2 to a cytoplasmic Cul3-based E3 ligase and suggest a model in which Keap1 coordinately regulates both Nrf2 accumulation and access to target genes.

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Taguchi, Keiko, and Masayuki Yamamoto. "The KEAP1–NRF2 System as a Molecular Target of Cancer Treatment." Cancers 13, no. 1 (December 26, 2020): 46. http://dx.doi.org/10.3390/cancers13010046.

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The Kelch-like ECH-associated protein 1 (KEAP1)—Nuclear factor erythroid-derived 2-like 2 (encoded by the Nfe2l2 gene; NRF2) system attracts extensive interest from scientists in basic and clinical cancer research fields, as NRF2 exhibits activity as both an oncogene and tumor suppressor, depending on the context. Especially unique and malignant, NRF2-addicted cancers exhibit high levels of NRF2 expression. Somatic mutations identified in the NRF2 or KEAP1 genes of NRF2-addicted cancers cause the stabilization and accumulation of NRF2. NRF2-addicted cancers hijack the intrinsic roles that NRF2 plays in cytoprotection, including antioxidative and anti-electrophilic responses, as well as metabolic reprogramming, and acquire a marked advantage to survive under severe and limited microenvironments. Therefore, NRF2 inhibitors are expected to have therapeutic effects in patients with NRF2-addicted cancers. In contrast, NRF2 activation in host immune cells exerts significant suppression of cancer cell growth, indicating that NRF2 inducers also have the potential to be therapeutics for cancers. Thus, the KEAP1–NRF2 system makes a broad range of contributions to both cancer development and suppression. These observations thus demonstrate that both NRF2 inhibitors and inducers are useful for the treatment of cancers with high NRF2 activity.

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Sun, Zheng, Shirley Zhang, Jefferson Y. Chan, and Donna D. Zhang. "Keap1 Controls Postinduction Repression of the Nrf2-Mediated Antioxidant Response by Escorting Nuclear Export of Nrf2." Molecular and Cellular Biology 27, no. 18 (July 16, 2007): 6334–49. http://dx.doi.org/10.1128/mcb.00630-07.

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ABSTRACT The transcription factor Nrf2 regulates cellular redox homeostasis. Under basal conditions, Keap1 recruits Nrf2 into the Cul3-containing E3 ubiquitin ligase complex for ubiquitin conjugation and subsequent proteasomal degradation. Oxidative stress triggers activation of Nrf2 through inhibition of E3 ubiquitin ligase activity, resulting in increased levels of Nrf2 and transcriptional activation of Nrf2-dependent genes. In this study, we identify Keap1 as a key postinduction repressor of Nrf2 and demonstrate that a nuclear export sequence (NES) in Keap1 is required for termination of Nrf2-antioxidant response element (ARE) signaling by escorting nuclear export of Nrf2. We provide evidence that ubiquitination of Nrf2 is carried out in the cytosol. Furthermore, we show that Keap1 nuclear translocation is independent of Nrf2 and the Nrf2-Keap1 complex does not bind the ARE. Collectively, our results suggest the following mechanism of postinduction repression: upon recovery of cellular redox homeostasis, Keap1 translocates into the nucleus to dissociate Nrf2 from the ARE. The Nrf2-Keap1 complex is then transported out of the nucleus by the NES in Keap1. Once in the cytoplasm, the Keap1-Nrf2 complex associates with the E3 ubiquitin ligase, resulting in degradation of Nrf2 and termination of the Nrf2 signaling pathway. Hence, postinduction repression of the Nrf2-mediated antioxidant response is controlled by the nuclear export function of Keap1 in alliance with the cytoplasmic ubiquitination and degradation machinery.

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Sekine, Hiroki, Keito Okazaki, Nao Ota, Hiroki Shima, Yasutake Katoh, Norio Suzuki, Kazuhiko Igarashi, Mitsuhiro Ito, Hozumi Motohashi, and Masayuki Yamamoto. "The Mediator Subunit MED16 Transduces NRF2-Activating Signals into Antioxidant Gene Expression." Molecular and Cellular Biology 36, no. 3 (November 16, 2015): 407–20. http://dx.doi.org/10.1128/mcb.00785-15.

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The KEAP1-NRF2 system plays a central role in cytoprotection. NRF2 is stabilized in response to electrophiles and activates transcription of antioxidant genes. Although robust induction of NRF2 target genes confers resistance to oxidative insults, how NRF2 triggers transcriptional activation after binding to DNA has not been elucidated. To decipher the molecular mechanisms underlying NRF2-dependent transcriptional activation, we purified the NRF2 nuclear protein complex and identified the Mediator subunits as NRF2 cofactors. Among them, MED16 directly associated with NRF2. Disruption ofMed16significantly attenuated the electrophile-induced expression of NRF2 target genes but did not affect hypoxia-induced gene expression, suggesting a specific requirement for MED16 in NRF2-dependent transcription. Importantly, we found that 75% of NRF2-activated genes exhibited blunted inductions by electrophiles inMed16-deficient cells compared to wild-type cells, which strongly argues that MED16 is a major contributor supporting NRF2-dependent transcriptional activation. NRF2-dependent phosphorylation of the RNA polymerase II C-terminal domain was absent inMed16-deficient cells, suggesting that MED16 serves as a conduit to transmit NRF2-activating signals to RNA polymerase II. MED16 indeed turned out to be essential for cytoprotection against oxidative insults. Thus, the KEAP1-NRF2-MED16 axis has emerged as a new regulatory pathway mediating the antioxidant response through the robust activation of NRF2 target genes.

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Türei, Dénes, Diána Papp, Dávid Fazekas, László Földvári-Nagy, Dezső Módos, Katalin Lenti, Péter Csermely, and Tamás Korcsmáros. "NRF2-ome: An Integrated Web Resource to Discover Protein Interaction and Regulatory Networks of NRF2." Oxidative Medicine and Cellular Longevity 2013 (2013): 1–9. http://dx.doi.org/10.1155/2013/737591.

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NRF2 is the master transcriptional regulator of oxidative and xenobiotic stress responses. NRF2 has important roles in carcinogenesis, inflammation, and neurodegenerative diseases. We developed an online resource, NRF2-ome, to provide an integrated and systems-level database for NRF2. The database contains manually curated and predicted interactions of NRF2 as well as data from external interaction databases. We integrated NRF2 interactome with NRF2 target genes, NRF2 regulating TFs, and miRNAs. We connected NRF2-ome to signaling pathways to allow mapping upstream NRF2 regulatory components that could directly or indirectly influence NRF2 activity totaling 35,967 protein-protein and signaling interactions. The user-friendly website allows researchers without computational background to search, browse, and download the database. The database can be downloaded in SQL, CSV, BioPAX, SBML, PSI-MI, and in a Cytoscape CYS file formats. We illustrated the applicability of the website by suggesting a posttranscriptional negative feedback of NRF2 by MAFG protein and raised the possibility of a connection between NRF2 and the JAK/STAT pathway through STAT1 and STAT3. NRF2-ome can also be used as an evaluation tool to help researchers and drug developers to understand the hidden regulatory mechanisms in the complex network of NRF2.

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McGrath-Morrow, Sharon, Thomas Lauer, Min Yee, Enid Neptune, Megan Podowski, Rajesh K. Thimmulappa, Michael O'Reilly, and Shyam Biswal. "Nrf2 increases survival and attenuates alveolar growth inhibition in neonatal mice exposed to hyperoxia." American Journal of Physiology-Lung Cellular and Molecular Physiology 296, no. 4 (April 2009): L565—L573. http://dx.doi.org/10.1152/ajplung.90487.2008.

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Increased oxidative stress is associated with perinatal asphyxia and respiratory distress in the newborn period. Induction of nuclear factor erythroid 2 p45-related factor (Nrf2) has been shown to decrease oxidative stress through the regulation of specific gene pathways. We hypothesized that Nrf2 attenuates mortality and alveolar growth inhibition in newborn mice exposed to hyperoxia. Nrf2+/+ and Nrf2−/− newborn mice were exposed to hyperoxia at 24 h. Survival was significantly less in Nrf2−/− mice exposed to 72 h of hyperoxia and returned to room air ( P < 0.0001) and in Nrf2−/− mice exposed to hyperoxia for 8 continuous days ( P < 0.005). To determine the response of Nrf2 target genes to hyperoxia, glutathione peroxidase 2 (Gpx2) and NAD(P)H:quinone oxidoreductase (NQO1) expression was measured from lung of newborn mice using real-time PCR. In the Nrf2+/+ mice, significant induction of lung Gpx2 and NQO1 above room air controls was found with hyperoxia. In contrast, Nrf2−/− mice had minimal induction of lung Gpx2 and NQO1 with hyperoxia. Expression of p21 and IL-6, genes not regulated by Nrf2, were also measured. IL-6 expression in Nrf2−/− lung was markedly induced by 72 h of hyperoxia in contrast to the Nrf2+/+ mice. p21 was induced in both Nrf2+/+ and Nrf2−/− lung by hyperoxia. Mean linear intercept (MLI) and mean chord length (MCL) were significantly increased in 14-day-old Nrf2−/− mice previously exposed to hyperoxia compared with Nrf2+/+ mice. The percentage of surfactant protein C (Sp-c+) type 2 alveolar cells in 14-day-old Nrf2−/− mice exposed to neonatal hyperoxia was also significantly less than Nrf2+/+ mice ( P < 0.02). In summary, these findings indicate that Nrf2 increases survival in newborn mice exposed to hyperoxia and that Nrf2 may help attenuate alveolar growth inhibition caused by hyperoxia exposure.

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Potteti, Haranatha R., Patrick M. Noone, Chandramohan R. Tamatam, Aparna Ankireddy, Sanjeev Noel, Hamid Rabb та Sekhar P. Reddy. "Nrf2 mediates hypoxia-inducible HIF1α activation in kidney tubular epithelial cells". American Journal of Physiology-Renal Physiology 320, № 3 (1 березня 2021): F464—F474. http://dx.doi.org/10.1152/ajprenal.00501.2020.

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Nuclear factor erythroid 2-related factor 2 (Nrf2) and hypoxia-inducible factor-1α (HIF1α) transcription factors protect against ischemic acute kidney injury (AKI) by upregulating metabolic and cytoprotective gene expression. In this study, we tested the hypothesis that Nrf2 is required for HIF1α-mediated hypoxic responses using Nrf2-sufficient (wild-type) and Nrf2-deficient ( Nrf2–/–) primary murine renal/kidney tubular epithelial cells (RTECs) and human immortalized tubular epithelial cells (HK2 cells) with HIF1 inhibition and activation. The HIF1 pathway inhibitor digoxin blocked hypoxia-stimulated HIF1α activation and heme oxygenase ( HMOX1) expression in HK2 cells. Hypoxia-mimicking cobalt (II) chloride-stimulated HMOX1 expression was significantly lower in Nrf2–/– RTECs than in wild-type counterparts. Similarly, hypoxia-stimulated HIF1α-dependent metabolic gene expression was markedly impaired in Nrf2–/– RTECs. Nrf2 deficiency impaired hypoxia-induced HIF1α stabilization independent of increased prolyl 4-hydroxylase gene expression. We found decreased HIF1α mRNA levels in Nrf2–/– RTECs under both normoxia and hypoxia-reoxygenation conditions. In silico analysis and chromatin immunoprecipitation assays demonstrated Nrf2 binding to the HIF1α promoter in normoxia, but its binding decreased in hypoxia-exposed HK2 cells. However, Nrf2 binding at the HIF1α promoter was enriched following reoxygenation, demonstrating that Nrf2 maintains constitutive HIF1α expression. Consistent with this result, we found decreased levels of Nrf2 in hypoxia and that were restored following reoxygenation. Inhibition of mitochondrial complex I prevented hypoxia-induced Nrf2 downregulation and also increased basal Nrf2 levels. These results demonstrate a crucial role for Nrf2 in optimal HIF1α activation in hypoxia and that mitochondrial signaling downregulates Nrf2 levels in hypoxia, whereas reoxygenation restores it. Nrf2 and HIF1α interact to provide optimal metabolic and cytoprotective responses in ischemic AKI.

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